SPATIAL STRUCTURE OPTIMIZATION AND PERFORMANCE RESEARCH OF COMPLEX FUNCTIONAL CHIMERIC FIBROUS CORPUSCLE

Du Jiliang; Qi Qi; Liu Han; Wang Xuxin; Wan Ping; Tian Shen

doi:10.19912/j.0254-0096.tynxb.2020-1060

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Acta Energiae Solaris Sinica ›› 2022, Vol. 43 ›› Issue (6) : 246-250. DOI: 10.19912/j.0254-0096.tynxb.2020-1060

SPATIAL STRUCTURE OPTIMIZATION AND PERFORMANCE RESEARCH OF COMPLEX FUNCTIONAL CHIMERIC FIBROUS CORPUSCLE

Du Jiliang^1,2, Qi Qi², Liu Han², Wang Xuxin², Wan Ping², Tian Shen^1,2

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Abstract

In this study, four different heterozygous fibrin scaffold proteins were successfully constructed. Cellulase and its extracellular assembly into a different spatial structure of chimeric designer cellulosome. The thermal stability and enzymatic hydrolysis kinetics of chimeric designer cellulosome were analyzed. The influence of different structures on the hydrolysis activity of designer cellosome and its optimal scaffold proteins structure was explored. Based on the Saccharomyces cerevisiae eby100 surface display system, the optimized scaffold protein was anchored and the performance of simultaneous saccharification and ethanol fermentation were assayed on PSAC. The results show that the enzyme activity of the designer cellosome remains relatively stable at 50 ℃ for 120 hours, the V_max=0.147 mg/（mL·min）, K_m=6.085 mg/mL and the reducing sugar production of ScafI-IV designer cellosome are all higher than those of the other cellosomes structural, which indicates the ScafI-IV type has a excellence substrate affinity and hydrolysis property. The maximum ethanol production is 1.12 g/L after 96 h, and the output is 0.263 g/g, which is equivalent to 51.50% of the theoretical value of ethanol yield. This result proves that the enzymolysis performance of the designer cellosome can be improved by structure optimizing of the scaffold proteins, which has reference significance for the research of artificially designing the cellosome.

Key words

Saccharomyces cerevisiae / cellulosic ethanol / spatial structural / surface display / fibrous corpuscle

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Du Jiliang, Qi Qi, Liu Han, Wang Xuxin, Wan Ping, Tian Shen. SPATIAL STRUCTURE OPTIMIZATION AND PERFORMANCE RESEARCH OF COMPLEX FUNCTIONAL CHIMERIC FIBROUS CORPUSCLE[J]. Acta Energiae Solaris Sinica. 2022, 43(6): 246-250 https://doi.org/10.19912/j.0254-0096.tynxb.2020-1060

References

[1] 王金兰, 王禄山, 刘巍峰, 等. 降解纤维素的“超分子机器”研究进展[J]. 生物化学与生物物理进展, 2011, 38(1): 28-35.
WANG J L, WANG L S, LIU W F, et al.Research advances on the assembly mode of cellulosomal macromolecular complexes[J]. Progress in biochemistry and biophysics, 2011, 38(1): 28-35.
[2] ARTZI L, BAYER E A, MORAÏS S. Cellulosomes: Bacterial nanomachines for dismantling plant polysaccharides[J]. Nature reviews microbiology, 2017, 15(2): 83-95.
[3] KATAEVA I A, UVERSKY V.Cellulosome (molecular anatomy and physiology of proteinaceous machines)[M]. Beijing: Chemical Industry Press, 2011.
[4] FIEROBE H P, MECHALY A, TARDIF C, et al.Design and production of active cellulosome chimeras—Selective incorporation of dockerin-containing enzymes into defined functional complexes[J]. Journal of biological chemistry, 2001, 276(24): 21257-21261.
[5] KOUKIEKOLO R, CHO H Y, AKIHIKO K, et al.Degradation of corn fiber by Clostridium cellulovorans cellulases and hemicellulases and contribution of scaffoldin protein CbpA[J]. Applied and environmental microbiology, 2005, 71(7): 3504-3511.
[6] MCCLENDON S D, MAO Z, SHIN H D, et al.Designer xylanosomes: Protein nanostructures for enhanced xylan hydrolysis[J]. Applied biochemistry and biotechnology, 2012, 167(3): 395-411.
[7] SUN J, WEN F, SI T, et al.Direct conversion of xylan to ethanol by recombinant Saccharomyces cerevisiae strains displaying an engineered minihemicellulosome[J]. Applied and environmental microbiology, 2012, 78(11): 13837-3845.
[8] TANG H, WANG J, WANG S, et al.Efficient yeast surface-display of novel complex synthetic cellulosomes[J]. Microbial cell factories, 2018, 17(1): 2-13.
[9] TIAN S, DU J L, BAI Z S, et al.Design and construction of synthetic cellulosome with three adaptor scaffoldins for cellulosic ethanol production from steam-exploded corn stover[J]. Cellulose, 2019, 26(15): 8401-8415.
[10] 陈宁, 杜济良, 田沈, 等. 自组装嵌合纤维小体酿酒酵母菌群的乙醇发酵研究[J]. 太阳能学报, 2018, 39(8): 2103-2109.
CHEN N, DU J L, TIAN S, et al.Study of ethanol fermentation of self-assembled designer cellulosome on Saccharomyces cerevisiae[J]. Acta energiae solaris sinica, 2018, 39(8): 2103-2109.
[11] ZHANG P, WANG B, XIAO Q, et al.A kinetics modeling study on the inhibition of glucose on cellulosome of Clostridium thermocellum[J]. Bioresource technology, 2015, 190: 36-43.
[12] MO C L, CHEN N, LYU T, et al.Direct ethanol production from steam-exploded corn stover using a synthetic diploid cellulase-displaying yeast consortium[J]. Bioresources, 2015, 10(3): 4460-4472.
[13] 王博伟, 闫雪晴, 何贤, 等. 纤维素可及性对木质纤维酶水解影响的研究进展[J]. 纤维素科学与技术, 2020, 28(1): 61-68.
WANG B W, YAN X Q, HE X, et al.Progress of the effects of cellulose accessibility on enzymatic hydrolysis of lignocellulose[J]. Journal of cellulose science and technology, 2020, 28(1): 61-68.
[14] STERN J, KAHN A, VAZANA Y, et al.Significance of relative position of cellulases in designer cellulosomes for optimized cellulolysis[J]. PLOS one, 2015, 10(5): 0127326.
[15] BARTH A, HENDRIX J, FRIED D, et al.Dynamic interactions type I cohesin modules fine-tune the structure of the cellulosome of Clostridium thermocellum[J]. Proceedings of the national academy of sciences of the United States of America, 2018, 115(48): E11274-E11283.